Rapid Quantification of Proteins in Complex Matrices using the DeNovix DS11 Microvolume Spectrophotometer

Recent advances in microvolume spectrometry have made significant impact on the workflow in life science laboratories by reducing the number of steps required in order to gain reliable data. Microvolume spectrophotometers require between 1-2 μL of sample and give highly accurate measurements in less than 20 seconds. During measurement, a droplet of sample is placed on a pedestal. An optical cell is formed when a top piece completing the light source assembly is placed onto the droplet and a column of fixed length is formed.

This application has become the gold standard in nucleic acids research labs but has significant limitations for characterisation and quantification of complex protein samples due to frequent column breakage. These limitations are due to sample properties such as surface tension and viscosity. Until now, there has been little confidence in this technology for protein applications.

In this poster, we will introduce the DeNovix DS-11 as the next generation in microvolume spectrophotometry. It’s patent pending SmartPath technology and proprietary alogorithms ensure that a column is formed every time prior to measurement ensuring ultimate confidence in the quality of results for the first time for all nucleic acid and protein applications.In addition, it offers the widest dynamic range eliminating the need for sample dilution prior to measurement as previously required. We will also discuss the ground breaking EasyApps® software suite which comes as standard on the custom AndroidTM tablet device, eliminating the need for an external PC. The small footprint of the DS-11 microvolume spectrophotometer (20 x 33 cm) makes it ideal for use in a busy multi-user environment, saving valuable bench space.

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Plasmodium falciparum is evolving resistance to Artemisinin Combination Therapy. The gene with the strongest association with resistance is K13. K13 is an ortholog of the well characterized transcriptional regulator Keap1. In this work we transcriptionally characterized a mutant with a transposon inserted in the K13 promoter region which results in dysregulation of K13 at 2 points of the intraerythrocytic cycle of the life-cycle to identify the processes regulated by K13.

PfABCk2 gene is ligated into pET21a+ vector with His-tag at C-terminus and transformed into BL-21 (DE3) competent cells that are verified through Miniprep and DNA sequencing. Furthermore, this gene construct is utilized to heterologous express this protein with IPTG and afterwards purified the recombinant protein kinase using nickel affinity chromatography as shown on 10% SDS-PAGE with the expected 36 kDa protein band.